Interleukin 6 (IL-6), also known as interferon-beta 2, 26 kDa protein and B cell stimulatory factor-2 (BSF-2), is a pleiotropic α-helical cytokine that plays important roles in acute phase reactions, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6 activity is essential for the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. It is secreted by multiple cell types as a 22 kDa - 28 kDa phosphorylated and variably glycosylated molecule. Mature human IL-6 is 183 amino acids (aa) in length and shares 41% aa sequence identity with mouse and rat IL-6. Alternate splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties.
IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R, triggering IL-6 R association with gp130 and gp130 dimerization. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R are generated by both alternate splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R elicit responses from gp130-expressing cells that lack cell surface IL-6 R. Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 R is predominantly restricted to hepatocytes, leukocytes, and lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R but not from other cytokines that utilize gp130 as a coreceptor.
原厂资料:
Product Summary:
The Quantikine Rat IL-6 Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-6 in cell culture supernates, rat serum, and plasma. It contains E. coli-expressed recombinant rat IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant protein. Results obtained using natural rat IL-6 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural rat IL-6.
Precision:
Intra-Assay Precision (Precision within an assay)Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
22
27
28
Mean
120
256
1,028
134
268
987
Standard Deviation
10.6
22.4
46.2
13.4
21.7
69.2
CV%
8.8
8.8
4.5
10
8.1
7
Recovery:
The recovery of rat IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Cell Culture Supernates (n=5)
103
85-118
Serum (n=5)
97
91-105
EDTA Plasma (n=5)
96
81-111
Linearity:
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of rat IL-6 in each matrix were diluted with Calibrator Diluent and then assayed.